SUMO-1 Recombinant Rabbit Monoclonal Antibody [SJ20-03]
cat.: ET1606-53
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IF-Tissue, IHC-P, IP, FC, ChIP
Clonality: Monoclonal
Clone number: SJ20-03
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 12 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human Human SUMO-1 aa 51-100 / 101.
Positive control: HeLa cell lysate, A431 cell lysate, MCF7 cell lysate, SH-SY5Y cell lysate, Jurkat cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, A431, NIH/3T3, human thyroid tissue, mouse trachea tissue, mouse esophagus tissue, mouse testis tissue, rat testis tissue, rat esophagus tissue.
Subcellular location: Nucleus membrane, Cytoplasm, Nucleus, Cell membrane.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
  FC
  IP
  ChIP
1:1,000-1:2,000
1:500
1:50-1:200
1:200-1:1,000
1:1,000
Use at an assay dependent concentration.
Use 0.5~2 μg for 25 μg of chromatin.
Uniprot #: SwissProt: P63165 Human | P63166 Mouse | Q5I0H3 Rat
Alternative names: DAP1 GAP modifying protein 1 GAP-modifying protein 1 GMP 1 GMP1 OFC10 PIC 1 PIC1 SENP2 Sentrin 1 Sentrin Small ubiquitin related modifier 1 Small ubiquitin-like modifier 1 Small ubiquitin-related modifier 1 SMT3 SMT3 homolog 3 SMT3 suppressor of mif two 3 homolog 1 SMT3, yeast, homolog 3 Smt3C SMT3H3 SUMO-1 SUMO1 SUMO1_HUMAN Ubiquitin homology domain protein PIC1 Ubiquitin Like 1 Ubiquitin like protein SMT3C Ubiquitin like protein UBL1 Ubiquitin-homology domain protein PIC1 Ubiquitin-like protein SMT3C Ubiquitin-like protein UBL1 UBL 1 UBL1
Images
ET1606-53_1.jpg Fig1: Western blot analysis of SUMO-1 on different lysates with Rabbit anti-SUMO-1 antibody (ET1606-53) at 1/1,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: A431 cell lysate
Lane 3: MCF7 cell lysate
Lane 4: SH-SY5Y cell lysate
Lane 5: Jurkat cell lysate
Lane 6: NIH/3T3 cell lysate
Lane 7: PC-12 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 12 kDa
Observed band size: 17/80 kDa

Exposure time: 1 minute;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1606-53) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ET1606-53_2.jpg Fig2: Immunocytochemistry analysis of A431 cells labeling SUMO-1 with Rabbit anti-SUMO-1 antibody (ET1606-53) at 1/500 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-SUMO-1 antibody (ET1606-53) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
ET1606-53_3.jpg Fig3: Immunocytochemistry analysis of NIH/3T3 cells labeling SUMO-1 with Rabbit anti-SUMO-1 antibody (ET1606-53) at 1/500 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-SUMO-1 antibody (ET1606-53) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
ET1606-53_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human thyroid tissue using anti-SUMO-1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1606-53, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1606-53_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded mouse trachea tissue with Rabbit anti-SUMO-1 antibody (ET1606-53) at 1/500 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1606-53) at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1606-53_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded mouse esophagus tissue with Rabbit anti-SUMO-1 antibody (ET1606-53) at 1/500 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1606-53) at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1606-53_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-SUMO-1 antibody (ET1606-53) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1606-53) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1606-53_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-SUMO-1 antibody (ET1606-53) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1606-53) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1606-53_9.jpg Fig9: Immunohistochemical analysis of paraffin-embedded rat esophagus tissue with Rabbit anti-SUMO-1 antibody (ET1606-53) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1606-53) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1606-53_10.jpg Fig10: Flow cytometric analysis of A431 cells labeling SUMO-1.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET1606-53, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
ET1606-53_11.jpg Fig11: Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and either SUMO-1 (ET1606-53) or Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.